CN102603769B - Sulfur-containing chromone compound and preparation method thereof and application in preparation of antitumor drugs - Google Patents

Abstract

The invention discloses a class of sulfur-containing chromone compounds, a preparation method thereof and an application in the preparation of antitumor drugs. The structure of sulfur-containing chromones is shown in formula (I), wherein compound 1: R ₁ =R ₂ =R ₃ =H; compound 2: R ₁ =R ₃ =H, R ₂ =CH ₃ ; compound 3 : R ₁ =R ₃ =Ac, R ₂ =H; Compound 4: R ₁ =R ₂ =R ₃ =Ac; Compound 5: R ₁ =R ₂ =Ac, R ₃ =H; Compound 6: R ₁ = Ac, R ₂ =R ₃ =H; Compound 7: R ₁ =(R)-MTPA, R ₂ =R ₃ =H; Compound 8: R ₁ =(S)-MTPA, R ₂ =R ₃ =H. Compound 1 and compound 2 were prepared and isolated from the fermentation broth of the fungus Penicillium oxalicum CCTCC AF 93256. Compound 1 was acetylated to obtain compound 3-6; compound 1 was subjected to Mosher esterification to obtain compounds 7 and 8. The sulfur-containing chromone compound of the present invention has the activity of inhibiting the growth of malignant melanoma cells, lung cancer cells, cervical cancer cells, breast cancer cells, laryngeal cancer cells, liver cancer cells, and intestinal cancer cells, so it has a good effect in the preparation of antitumor drugs. application prospects. Formula (I).

Description

A class of sulfur-containing chromone compounds, their preparation method and their application in the preparation of antitumor drugs

Technical field:

The invention belongs to the field of biotechnology, and in particular relates to a class of sulfur-containing chromone compounds, a preparation method thereof and an application in preparation of antitumor drugs.

Background technique:

Fungi can produce secondary metabolites such as alkaloids, peptides, polyketides, steroids, and terpenes, many of which have significant anti-tumor, anti-bacterial, anti-fungal, and anti-insect biological activities. Searching for active compounds from fungi is a hotspot in the development of natural medicines. Chromone compounds are a class of substances that exist in nature and have various biological activities. Whether they are naturally occurring or synthesized, they have attracted widespread attention from chemists and others. However, sulfur-containing chromone compounds are very rare.

Invention content:

The first object of the present invention is to provide a class of sulfur-containing chromone compounds with antitumor activity.

The present invention separates and obtains a new class of sulfur-containing chromone compounds from the fermentation broth of the fungus Penicillium oxalicum CCTCCAF 93256 through various column chromatography and one-dimensional and two-dimensional nuclear magnetic resonance spectra. This sulfur-containing chromone compound has The cytotoxic activity that inhibits the growth of various cancer cells can be used to prepare antitumor drugs, thereby achieving the purpose of the present invention.

The sulfur-containing chromone compound of the present invention has a structure as shown in formula (I):

Formula (I)

Compound 1: R ₁ =R ₂ =R ₃ =H; Compound 2: R ₁ =R ₃ =H, R ₂ =CH ₃ ; Compound 3: R ₁ =R ₃ =Ac, R ₂ =H; Compound 4 : R ₁ =R ₂ =R ₃ =Ac; Compound 5: R ₁ =R ₂ =Ac, R ₃ =H; Compound 6: R ₁ =Ac, R ₂ =R ₃ =H; Compound 7: R ₁ = (R)-MTPA, R ₂ =R ₃ =H; compound 8: R ₁ =(S)-MTPA, R ₂ =R ₃ =H.

The second object of the present invention is to provide a preparation method of the sulfur-containing chromones shown in formula (I).

The preparation method of the sulfur-containing chromone compound shown in formula (I) of the present invention is characterized in that it comprises the following steps:

(1) prepare the fermented liquid of fungus Penicillium oxalicum CCTCC AF 93256;

(2) Extract the fermentation broth obtained in step (1) with ethyl acetate, dichloromethane or chloroform solvent, and concentrate to obtain ethyl acetate extract, dichloromethane extract or chloroform extract;

(3) The ethyl acetate extract, dichloromethane extract or chloroform extract described in step (2) is subjected to normal pressure silica gel column chromatography, and the chloroform-methanol, chloroform-acetone, chloroform-ethyl acetate, petroleum Ether-acetone or sherwood oil-ethyl acetate solvent system is eluent, carries out gradient elution from volume ratio 100: 0 to 0: 100, uses thin-layer chromatography to track and combine component, will be able to be obtained on thin-layer chromatography The components developed with the chloroform-acetone solvent system with a volume ratio of 8:2 are subjected to gel column chromatography to obtain crude products, which are purified to obtain sulfur-containing chromone compounds 1 and 2 shown in formula (I);

(4) Compound 1 was dissolved in anhydrous pyridine and reacted with acetic anhydride at room temperature using 4-dimethylaminopyridine as a catalyst to prepare compound 3-6.

(5) Compound 1 was dissolved in anhydrous pyridine at room temperature with (S)-2-methoxy-2-trifluoromethylphenylacetyl chloride and (R)-2-methoxy-2-trifluoro Compounds 7 and 8 were prepared by reacting methylphenylacetyl chloride.

The fungus Penicillium oxalicum CCTCC AF 93256 described in step (1) was purchased from the China Center for Type Culture Collection (CCTCC), address: China, Wuhan, Wuhan University, and the preservation number is CCTCC AF 93256.

The fermentation broth of the fungus Penicillium oxalicum CCTCC AF 93256 described in the step (1) can be prepared by inoculating the fungus Penicillium oxalicum CCTCC AF 93256 into a suitable medium for fungi of the genus Penicillium under normal fermentation conditions. The preferred preparation method is to inoculate the fungus Penicillium oxalicum CCTCC AF 93256 in the PDA agar medium, and cultivate it at 28°C for 3 days to obtain a plate with cultured bacteria, then inoculate the bacteria in the plate into the PDA medium, and inoculate at a speed of 200r /min shaker, temperature 28 ℃ for 3 days, to obtain the seed liquid, then inoculate the seed liquid in the PDA medium, and cultivate it statically at room temperature for 30 days, to obtain the fermentation liquid, the PDA agar medium per liter contains potato 200g, glucose 20g, sea salt 30g, agar 20g, and the balance is water. The PDA medium contains 200g of potatoes per liter, glucose 20g, sea salt 30g, and the balance is water.

The extraction described in step (2) preferably uses ethyl acetate, and the described concentration can adopt conventional methods such as concentration under reduced pressure.

The purification described in step (3) can adopt chromatographic column separation or recrystallization.

Through in vitro anti-tumor activity screening experiments, the results show that the sulfur-containing chromone compounds shown in formula (I) of the present invention can inhibit malignant melanoma cell line A375, lung cancer cell line A549, cervical cancer cell line HeLa, breast cancer cell line The growth of cell line MCF-7, laryngeal cancer cell line Hep-2, liver cancer cell line HepG2, intestinal cancer cell line SW-620, their IC ₅₀ values are as shown in table 3, prove thus the present invention as formula (I ) The sulfur-containing chromone compounds shown in ) can be used to prepare antitumor drugs.

Therefore, the third object of the present invention is to provide the application of the sulfur-containing chromone compounds represented by formula (I) in the preparation of antitumor drugs.

Preferably, when the sulfur-containing chromone compound is Compound 1, the antineoplastic drug is an anti-melanoma, lung cancer, cervical cancer, breast cancer, laryngeal cancer, liver cancer or intestinal cancer drug.

Preferably, when the sulfur-containing chromone compound is compound 2, compound 3 or compound 4, the antineoplastic drug is an anti-melanoma, lung cancer, cervical cancer or intestinal cancer drug.

Preferably, when the sulfur-containing chromone compound is compound 5, the anti-tumor compound is an anti-melanoma, cervical cancer or intestinal cancer drug.

Preferably, when the sulfur-containing chromone compound is compound 6, the anti-tumor compound is an anti-melanoma or intestinal cancer drug.

Preferably, when the sulfur-containing chromone compound is compound 7, the antineoplastic drug is an anti-melanoma, lung cancer, cervical cancer or intestinal cancer drug.

An antineoplastic drug is characterized in that it comprises an effective amount of thiochromone compounds as active ingredients and a pharmaceutically acceptable carrier.

The sulfur-containing chromone compound shown in formula (I) of the present invention has a stronger effect on the growth of malignant melanoma cells, lung cancer cells, cervical cancer cells, breast cancer cells, laryngeal cancer cells, liver cancer cells, and intestinal cancer cells. It has good application prospect in the preparation of antitumor drugs.

Description of drawings:

Fig. 1 is the measured CD figure of compound 1 in dichloromethane;

Fig. 2 is the measured CD diagram of compound 1 in DMSO containing Mo ₂ (OAc) ₄ .

Detailed ways:

The following examples are to further illustrate the present invention, rather than limit the present invention.

Embodiment 1: Preparation of sulfur-containing chromone compounds 1 and 2

Each liter of PDA medium is prepared as follows: 200 grams of potatoes, 20 grams of glucose, and 30 grams of sea salt are mixed, and the volume is adjusted to 1L with water. PDA medium was filled into about 200 500mL Erlenmeyer flasks, each bottle was about 150mL, and sterilized by high-pressure steam at 121°C for 25 minutes.

Each liter of PDA agar medium is prepared as follows: mix 200 grams of potatoes, 20 grams of glucose, 30 grams of sea salt, and 20 grams of agar, and dilute to 1L with water. Autoclave at 121°C for 25 minutes and set aside.

Use a pipette gun to inoculate about 2 microliters of the fungus Penicillium oxalicum CCTCC AF 93256 into the PDA agar medium, and cultivate it at 28°C for 3 days to obtain a plate with cultured bacteria, and then pick about 3 microliters from the plate with a bamboo stick. Microliter strains were inoculated into the Erlenmeyer flask containing 150mL of the above-mentioned PDA medium, and cultivated for 3 days at a temperature of 28°C on a shaker (rotating speed 200r/min) to obtain a seed solution. Inoculate 7.5mL of seed liquid in the Erlenmeyer flask, and after static cultivation at room temperature (28°C) for 30 days, collect the fermentation liquid.

30 L of the fermentation liquid obtained by culturing in PDA medium was extracted with ethyl acetate, and concentrated under reduced pressure to obtain 16 g of ethyl acetate extract. After the ethyl acetate extract was mixed with normal-phase silica gel (100-200 mesh) by dry method, it was loaded into a glass chromatography column (containing about 300 g of normal-phase silica gel 100-200 mesh), and carried out normal temperature column chromatography, with chloroform-methanol As the eluent, gradient elution was performed from volume ratio 100:0 to 0:100, and the fractions were combined according to thin layer chromatography (GF ₂₅₄ silica gel plate), the elution solvent was recovered, and the fractions were evaporated to dryness and transferred with methanol. The component (2.4g) that can develop with the chloroform-acetone solvent system of volume ratio 8:2 on thin layer chromatography (2.4g) is through gel column chromatography (diameter 18mm, column length 1600mm, and gel is sephedex LH-20, Mobile phase is the chloroform-methanol of volume ratio 1: 1) separation, collects fraction, separates out crude product, and crude product adopts high performance liquid phase semi-preparative (detection wavelength is 254nm, and flow rate is 3mL/min, and chromatographic column is YMC-Pack 10mm * 250mm , the mobile phase was methanol-water with a volume ratio of 45:55) and compounds 1 and 2 were obtained by separation and purification at peak eluting times of 20 and 23 minutes, respectively.

The structural analysis of compound 1 is as follows:

High-resolution mass spectrometry (HRESIMS) gives quasi-molecular ion peaks at m/z 447.0693[M+Na] ⁺ , 425.0919[M+H] ⁺ , combined with NMR spectral data, it is known that the molecular formula of compound 1 is C ₁₉ H ₂₀ O ₉ S. ¹³ C spectrum and DEPT spectrum show that there are 19 carbon atoms in the molecule, including 3 methyl groups [δ _C 22.34, 53.02, 53.77], 2 methylene groups [δ _C 35.45, 37.96], 4 methine groups [ _δC 50.77, 71.21, 107.86, 113.04], and 10 quaternary carbons [ _δC 79.61, 109.12, 120.51, 147.22, 157.12, 160.88, 171.01, 173.16, 173.41, 178.87]. The hydrogen spectrum shows 2 benzene ring hydrogen signals [δ _H 6.63 (1H, s), 6.72 (1H, s)], 2 methoxy groups [δ _H 3.82 (3H, s), 3.88 (3H, s)], and a downfield phenolic hydroxyl active hydrogen [δ _H 12.6]. These NMR signals are similar to those of the basic skeletons of compounds (2'R)-aloesol and (2'S)-aloesol [1.Kashiwada, Y.; Nonaka, GI; Nishioka, I.Chem.Pharm.Bull.1984, 32, 3493-3500.2.Che, QM; Akao, T.; Hattori, M.; Kyoichi, K.; Structural features of the class of compounds (A ring and B ring basic skeleton).

In the HMBC spectrum, H-2 is related to C-1, C-3, C-4, C-8a, C-17, H-4 is related to C-2, C-3, C-3, C-4a, C-8a, C-17 is related, Me-17 is related to C-2, C-3, C-4, which shows that compound 1 has a 1,2,3,5-tetrasubstituted benzene ring (A ring).HMBC In the spectrum, H-7 is related to C-6, C-9, C-10, and in ^{the 1} H- ¹ H COZY spectrum, H-7 is related to H-6, and combined with H-7/H-6/C-7 The 1D NMR data of /C-6 speculated that there is a sulfur-containing five-membered ring (C ring) connected to the B ring. In addition, in the HMBC spectrum, H-6 is related to C-11, C-14, H-11 is related to C-6, C-12, C-13, Me-15 is related to C-14, Me-16 is related to C- 13 correlation, combined with the correlation of H-11 and H-12 in the ¹ H- ¹ H COZY spectrum, it is speculated that there is a 1,5-diformyl-2,4-dihydroxy-substituted methyl dipivalate fragment connected to the C ring. In addition, the absolute configurations of chiral carbon atoms C-6, C-11, and C-13 were further confirmed by NOESY spectrum, Mosher esterification reaction, and circular dichroism (see Figures 1, 2,) as S, S, R. Therefore, it is determined that the structure of compound 1 is shown in formula (I), and its R ₁ =R ₂ =R ₃ =H. Compound 1 is a yellow powder, easily soluble in chloroform, ethyl acetate, methanol, and DMSO, but poorly soluble in water. The specific rotation value [a] ²⁰ _D =+31.43 (c, 0.42in CH ₃ Cl).

The hydrogen spectrum and carbon spectrum data of compound 2 are very similar to those of compound 1, except that there is an extra methoxyl group. It is speculated that the methoxyl group is connected to the C-11 position through the NMR data of compound 1 and the relevant points in the HMBC spectrum. Therefore, the structure of compound 2 was confirmed as shown in formula (I), where R ₁ =R ₃ =H, R ₂ =CH ₃ .

The hydrogen spectrum and carbon spectrum data of compounds 1 and 2 are shown in Table 1 and Table 2.

Formula (I)

Compound 1: R ₁ =R ₂ =R ₃ =H; Compound 2: R ₁ =R ₃ =H, R ₂ =CH ₃ ; Compound 3: R ₁ =R ₃ =Ac, R ₂ =H; Compound 4 : R ₁ =R ₂ =R ₃ =Ac; Compound 5: R ₁ =R ₂ =Ac, R ₃ =H; Compound 6: R ₁ =Ac, R ₂ =R ₃ =H; Compound 7: R ₁ = (R)-MTPA, R ₂ =R ₃ =H; compound 8: R ₁ =(S)-MTPA, R ₂ =R ₃ =H.

Table 1. Proton spectrum NMR data of compound 1-6 (in CDCl ₃ solvent)

The carbon spectrum NMR data of table 2. compound 1-4 (in CDCl ₃ solvents)

Embodiment 2: Preparation of sulfur-containing chromone compounds 3-6

Weigh 4.0 mg of compound 1 and dissolve it in a 5 mL flask containing 0.8 mL of anhydrous pyridine, add 0.8 mL of acetic anhydride and 2.0 mg of 4-dimethylaminopyridine (as a catalyst) to the solution, and react at room temperature for 11 hours. After the solvent was evaporated to dryness under reduced pressure, the crude product was semi-preparatively prepared by high performance liquid phase (detection wavelength was 254nm, flow rate was 3mL/min, chromatographic column was YMC-Pack 10mm×250mm, mobile phase was CH ₃ CN/H ₂ O from volume Gradient elution from 20% to 100%, samples with peak times of 19.6 min, 21.4 min, 23.9 min, and 26.0 min were collected respectively to obtain compounds 3, 4, 5, and 6.

Compound 3-6 is an acetylated product of compound 1. According to the nuclear magnetic resonance data of compound 3-6, and compared with the nuclear magnetic resonance data of compounds 1 and 2, the structure of compound 3-6 is determined as shown in formula (I), Wherein compound 3: R ₁ =R ₃ =Ac, R ₂ =H; compound 4: R ₁ =R ₂ =R ₃ =Ac; compound 5: R ₁ =R ₂ =Ac, R ₃ =H; compound 6: R ₁ =Ac, R ₂ =R ₃ =H.

The hydrogen spectrum and carbon spectrum data of compound 3-6 are listed in Table 1 and Table 2.

Embodiment 3: Preparation of sulfur-containing chromone compounds 7 and 8

Weigh 2.0 mg of compound 1 and dissolve it in a 5 mL flask containing 0.5 mL of anhydrous pyridine, add Mosher reagent 10 μL of (S)-2-methoxy-2-trifluoromethylphenylacetyl chloride and 1.0 mg of DMAP to the solution (as a catalyst), react at room temperature for 9 hours. After the solvent was evaporated to dryness under reduced pressure, the crude product was semi-preparatively used in high performance liquid phase (detection wavelength is 254nm, flow rate is 3mL/min, chromatographic column is YMC-Pack 10mm×250mm, mobile phase is _CH3CN / _H2O (from 20% to 100% gradient elution, volume ratio v/v) separation and purification to obtain compound 7.

The method for preparing compound 8 is similar to that of compound 7, except that the Mosher reagent is replaced by (R)-2-methoxy-2-trifluoromethylphenylacetyl chloride.

Compounds 7 and 8 are Mosher esterification products of compound 1. According to the nuclear magnetic resonance data and comparing with the nuclear magnetic resonance data of compounds 1 and 2, the structures of compounds 7 and 8 are determined as shown in formula (I). Compound 7: R ₁ = (R)-MTPA, R ₂ =R ₃ =H; compound 8: R ₁ =(S)-MTPA, R ₂ =R ₃ =H.

The proton spectrum data of compounds 7 and 8 are as follows.

Compound 7: ¹ H NMR (500MHz, CDCl ₃ ) δ2.42 (3H, s, H-18), 2.80 (1H, dd, J=8.4, 17.3Hz, H-7a), 2.75 (1H, dd, J =8.3, 17.3Hz, H-7b), 3.03(1H, dd, J=4.1, 15.1Hz, H-12a), 2.98(1H, dd, J=7.1, 15.1Hz, H-12b), 3.82(3H , s, H-16), 3.88 (3H, s, H-17), 3.83 (1H, dd, J=8.0, 8.5Hz, H-6), 5.40 (1H, dd, J=4.5, 7.2Hz, H-13), 6.65 (1H, s, H-2), 6.74 (1H, s, H-4), 12.05 (OH-1).

Compound 8: ¹ H NMR (500MHz, CDCl ₃ ) δ2.41 (3H, s, H-18), 3.08 (1H, dd, J=8.5, 17.5Hz, H-7a), 3.22 (1H, dd, J =8.0, 17.5Hz, H-7b), 3.16(1H, dd, J=3.5, 15.0Hz, H-12a), 3.02(1H, dd, J=8.5, 15.0Hz, H-12b), 3.77(3H , s, H-17), 3.85 (3H, s, H-16), 3.94 (1H, dd, J=8.5, 8.0Hz, H-6), 5.40 (1H, dd, J=3.5, 8.5Hz, H-13), 6.65 (1H, s, H-2), 6.74 (1H, s, H-4), 12.05 (OH-1).

Example 4: In vitro anti-tumor activity screening experiments of sulfur-containing chromone compounds 1, 2, 3, 4, 5, 6 and 7 shown in formula (I):

Collect malignant melanoma cell line A375, lung cancer cell line A549, cervical cancer cell line HeLa, breast cancer cell line MCF-7, laryngeal cancer cell line Hep-2, liver cancer cell line HepG2 and intestinal cancer cell line in logarithmic growth phase SW-620 was suspended in 10% serum 1640 medium, then inoculated in 96-well culture plate, the number of cells per well was 5000/80 μL, and cultured in a 5% CO ₂ incubator at 37°C. Dilute thiochromone compounds (compounds 1, 2, 3, 4, 5, 6 or 7) with 10% serum 1640 medium to a concentration of 3.125 μg/mL, 6.25 μg/mL, 12.5 μg/mL, 25 μg /mL, 50μg/mL, 100μg/mL, 200μg/mL of the experimental solution. On the next day, 20 μL of the experimental solutions with concentrations of 3.125 μg/mL, 6.25 μg/mL, 12.5 μg/mL, 25 μg/mL, 50 μg/mL, 100 μg/mL and 200 μg/mL were added to the culture plates of different experimental groups, And make the final concentration in each well reach the test (concentration of the experimental group adopts double dilution). In addition, the same amount of 10% serum 1640 medium was added to the negative control group. After 48 hours, discard the culture medium in the experimental group and the control group, add 20 μL (2.5 mg/mL) of MTT to each well, continue to incubate for 4 hours, then add 100 μL of DMSO to each well to stop the reaction, place at 37°C for 20 minutes, and detect each well with a microplate reader. The absorbance A value of the hole at 570nm was used to calculate the cell growth inhibition rate. Cell growth rate=(experimental group OD÷control group OD)×100%.

Experimental results show that sulfur-containing chromone compounds 1-7 represented by formula (I) can inhibit malignant melanoma cell line A375, lung cancer cell line A549, cervical cancer cell line HeLa, breast cancer cell line MCF-7, laryngeal cancer The growth of cell line Hep-2, liver cancer cell line HepG2, and intestinal cancer cell line SW-620, and their IC50 values are shown in Table 3. Therefore, the sulfur-containing chromone compounds represented by the formula (I) of the present invention can be used in the preparation of antitumor drugs.

The cytotoxic activity of table 3 compound 1-7 to 7 kinds of tumor cells

Note: "-" means no test due to insufficient sample volume.

Claims (9)

1. Shown in formula I containing Thiochromone compound:

   

   Compound 1:R wherein ₁=R ₂=R ₃=H; Compound 2:R ₁=R ₃=H, R ₂=CH ₃; Compound 3:R ₁=R ₃=Ac, R ₂=H; Compound 4:R ₁=R ₂=R ₃=Ac; Compound 5:R ₁=R ₂=Ac, R ₃=H; Compound 6:R ₁=Ac, R ₂=R ₃=H; Compound 7:R ₁=(R)-MTPA, R ₂=R ₃=H; Compound 8:R ₁=(S)-MTPA, R ₂=R ₃=H.
2. 2. the preparation method containing Thiochromone compound claimed in claim 1, is characterized in that, comprises the following steps:

   (1) prepare the fermented liquid of fungi Penicillium oxalicum CCTCC AF 93256;

   (2) ethyl acetate, methylene dichloride or the chloroform solvent extraction for fermented liquid that step (1) are obtained, concentrated ethyl acetate extract, dichloromethane extract or the chloroform extract of obtaining;

   (3) by the described ethyl acetate extract of step (2), dichloromethane extract or chloroform extract process normal pressure silica gel column chromatography, take chloroform-methanol, chloroform-acetone, chloroform-ethyl acetate, sherwood oil-acetone or petroleum ether-ethyl acetate solvent systems is eluent, carry out gradient elution from volume ratio 100:0 to 0:100, follow the trail of and merge component with thin-layer chromatography, to on thin-layer chromatography, can use the component that chloroform-the acetone solvent system is launched of volume ratio 8:2, obtain crude product through gel filtration chromatography, purified, obtain compound 1 claimed in claim 1 and 2;

   (4) compound 1 is dissolved in anhydrous pyridine and reacts with acetic anhydride as catalyzer and prepare compound 3-6 claimed in claim 1 with DMAP in room temperature;

   (5) compound 1 is dissolved in anhydrous pyridine and prepares compound 7 claimed in claim 1 and 8 with (S)-2-methoxyl group-2-trifluoromethyl phenyllacetyl chloride and (R)-2-methoxyl group-2-trifluoromethylbenzene excess acetyl chloride respectively in room temperature;

   The preparation method of the fermented liquid of the fungi Penicillium oxalicum CCTCC AF 93256 described in step (1) is inoculated in fungi Penicillium oxalicum CCTCC AF 93256 in the PDA nutrient agar, cultivate 3 days for 28 ℃, obtain cultivating the flat board that bacterial classification is arranged, then bacterial classification in flat board is inoculated in the PDA substratum, in rotating speed 200r/min shaking table, 28 ℃ of cultivations of temperature 3 days, obtain seed liquor, seed liquor is inoculated in the PDA substratum again, in the standing cultivation of room temperature 30 days, obtain fermented liquid, every liter of described PDA nutrient agar contains potato 200g, glucose 20g, sea salt 30g, agar 20g, surplus is water, every liter of described PDA substratum contains potato 200g, glucose 20g, sea salt 30g, surplus is water, the described extraction ethyl acetate of step (2), described concentrated employing concentrating under reduced pressure, the described purifying of step (3) adopts chromatographic column to separate or recrystallization.
3. 3. claimed in claim 1 containing the application of Thiochromone compound in preparing antitumor drug.
4. 4. application according to claim 3, is characterized in that, described is compound 1 claimed in claim 1 containing Thiochromone compound, and described antitumor drug is melanoma, lung cancer, cervical cancer, mammary cancer, laryngocarcinoma, liver cancer or bowelcancer medicine.
5. 5. application according to claim 3, is characterized in that, described is compound 2 claimed in claim 1, compound 3 or compound 4 containing Thiochromone compound, and described antitumor drug is melanoma, lung cancer, cervical cancer or bowelcancer medicine.
6. 6. application according to claim 3, is characterized in that, described is compound 5 claimed in claim 1 containing Thiochromone compound, and described antitumor drug is melanoma, cervical cancer or bowelcancer medicine.
7. 7. application according to claim 3, is characterized in that, described is compound 6 claimed in claim 1 containing Thiochromone compound, and described antitumor drug is melanoma or bowelcancer medicine.
8. 8. application according to claim 3, is characterized in that, described is compound 7 claimed in claim 1 containing Thiochromone compound, and described antitumor drug is melanoma, lung cancer, cervical cancer or bowelcancer medicine.
9. 9. an antitumor drug, is characterized in that, comprises claimed in claim 1 containing Thiochromone compound and acceptable carrier pharmaceutically as activeconstituents of significant quantity.

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